MANIPULATION OF AN ALTERNATIVE RESPIRATORY PATHWAY IN PHOTO-AUTOTROPHS
First Claim
1. A method for increasing TAG production in an algal cell during imbalanced growth conditions, the method comprising:
- knocking out an AOX gene.
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Abstract
Exemplary methods for increasing TAG production in an algal cell during imbalanced growth conditions are provided. Some methods comprise knocking out an AOX gene, wherein the AOX gene produces an amino acid sequence having substantial similarity to the amino acid sequence of SEQ. ID. NO. 2. In further methods, the algal cell may be of genus Nannochloropsis. The AOX gene may be replaced by a construct having a nucleotide sequence having substantial similarity to SEQ ID. NOS. 3 through 5 (inclusive), wherein each of the sequences are next to or in close proximity to one another in a linear fashion. In some methods, the AOX gene may be replaced via homologous recombination. As a result, lipid production by the selected recombinant algal cell may be increased over that produced by a wild-type algal cell.
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Citations
19 Claims
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1. A method for increasing TAG production in an algal cell during imbalanced growth conditions, the method comprising:
knocking out an AOX gene. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
- 13. A transformation construct, the transformation construct having a first sequence of DNA similar to a corresponding first sequence of nuclear DNA of an algal cell, the transformation construct having a second sequence of DNA similar to a corresponding second sequence of nuclear DNA of the algal cell, and the transformation construct having a sequence of DNA of interest inserted between the first and second sequences of the transformation construct.
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19. A transformation method for introducing deoxyribonucleic acid (DNA) into the nucleus of an algal cell, the method comprising:
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preparing a transformation construct, the transformation construct having a first sequence of DNA similar to a corresponding first sequence of nuclear DNA, the transformation construct having a second sequence of DNA similar to a corresponding second sequence of the nuclear DNA, the transformation construct having a sequence of DNA of interest inserted between the first and second sequences of DNA of the transformation construct, and transforming a target sequence of DNA inserted between the first and second corresponding sequences of the nuclear DNA, resulting in replacement of the target sequence of DNA with the sequence of DNA of interest, wherein the target sequence has substantial similarity to SEQ. ID. NO. 1.
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Specification